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. 2018 Feb 10;23(2):381.
doi: 10.3390/molecules23020381.

Separation and Purification of Fructo-Oligosaccharide by High-Speed Counter-Current Chromatography Coupled with Precolumn Derivatization

Wenjuan Duan  1 , Wenhua Ji  2 , Yuanan Wei  3 , Ruixuan Zhao  4 , Zijian Chen  5 , Yanling Geng  6 , Feng Jing  7 , Xiao Wang  8
Affiliations

Separation and Purification of Fructo-Oligosaccharide by High-Speed Counter-Current Chromatography Coupled with Precolumn Derivatization

Wenjuan Duan et al. Molecules. .

Abstract

High-speed counter-current chromatography (HSCCC) coupled with precolumn derivatization was developed for isolating and purifying fructo-oligosaccharides (FOSs). Firstly, the total FOSs were precolumn derivatized and then separated by high-speed counter-current chromatography (HSCCC) with two-phase solvent system petroleum ether-n-butanol-methanol-water (3:2:1:4, v/v). Secondly, the obtained compounds were deacetylated and the fructo-oligosaccharides (FOSs) with high purity were obtained. Their structures were identified by mass spectrometry (MS) and nuclear magnetic resonance (NMR). This research successfully established a novel strategy for separation and purification of FOS. There is no doubt that the application of the research will be beneficial for the quantitative and qualitative analysis of products containing FOSs.

Keywords: fructo-oligosaccharide; high-speed counter-current chromatography; precolumn derivatization; separation and purification.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Roadmap of separation of FOSs (a) total FOSs; (b) the structure of kestose; (c) the structure of 1,1-kestotetraose; (d) the structure of 1,1,1-kestopentaose.
Figure 2
Figure 2
HPLC chromatograms of total acetylated FOSs and separated compounds: (a) HPLC chromatograms of total acetylated FOSs; (b) HPLC chromatograms of compound I; (c) HPLC chromatograms of compound II; (d) HPLC chromatograms of HSCCC peak of compound III.
Figure 3
Figure 3
HSCCC chromatogram of the acetylated fructo-oligosaccharides. (a) Solvent system: petroleum ether–ethyl acetate–methanol–water (1:1:1:1, v/v); stationary phase: upper phase; mobile phase: lower phase; flow rate: 2.0 mL/min; revolution speed: 800 rpm; retention of stationary phase: 57.0%; sample load: 200 mg; detection: ELSD; (b) Solvent system: petroleum ether–n-butanol–methanol–water (3:2:1:4, v/v); stationary phase: upper phase; mobile phase: lower phase; flow rate: 2.0 mL/min; revolution speed: 800 rpm; retention of stationary phase: 53.0%; sample load: 200 mg; detection: ELSD.
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